摘要
目的:应用定量荧光PCR方法检测21-三体综合征及性染色体数目异常疾病,通过优化多重定量荧光PCR体系,建立一种快速简便的染色体异常疾病的检测方法。方法:针对21号染色体上特异的3个STR位点(D21S1435、D21S1411、D21S11),X染色体上的两个STR位点(DXS981、DXS6809),X、Y染色体上共有的STR位点X22及性别特异性位点AMXY设计引物,并在引物的5'端标记荧光,对202例外周血标本提取基因组DNA进行七重定量荧光PCR扩增,使用ABI310测序仪进行PCR结果分析,根据国际统一判断标准进行结果判断。结果:QF-PCR分析202例标本中97例为21-三体标本,62例为性染色体异常标本,其中96例21-三体标本(包括1例易位型和1例嵌合型)和56例性染色体异常标本的诊断结果与染色体核型分析结果一致,QF-PCR检测21-三体和性染色体异常灵敏度和特异度分别为90.3%,98%,96%,99.1%。结论:七重定量荧光PCR诊断结果与染色体核型诊断结果具有同一性;QF-PCR在产前诊断染色体异常疾病灵敏度高、特异性强,具有较高实用价值和市场前景。
Objective： To detect the 21 - trisomy syndrome and hererosome numerical abnormality disease using quanti- tative fluorescent polymerase chain reaction （QF- PCR）, and to set a rapid and reliable detection for chromosomal ab- normality disease based on the optimization of multiple QF -PCR. Methods： Specific primers were designed and synthesized according to the 3 STR sites （D21S1435, D21Sll and D21 S1411 ） on chromosome 21,2 STR sites （ DXS981 and DXS6809 ） on chromosome X, the STR site （X22） co -located on chromosome X and Y, and the gender specific site AMXY. And the primers were labelled （5＇end） with a fluorescent dye （ FAM or HEX）. DNA samples were extracted from peripheral blood of 202 cases and amplified using QF - PCR, and then the amplification products were analyzed by ABI 310 sequenator and evaluated under the international unified standard. Re- suits： QF -PCR analysis showed that 97 out of 202 cases were identified as trisomy -21, and 62 cases as heterosome abnor- mality, among which the diagnosis of 96 cases with trisomy -21 （including a case of translocation and a case of mosaicism） and 56 cases with heterosome abnormality were coincident with chromosome karyotype analysis. The sensitivity and specificity of QF - PCR in the detection of 21 - trisomy syndrome and heterosome abnormality were （90.3%, 98% ） and （96%, 99. 1% ）, respectively. Conclusion： The diagnosis using QF - PCR or chromosome karyotype analysis show high concordance. And the application of QF - PCR in prenatal diagnosis will make higher practical values and better market prospects for its high sensitivity and strong specificity.
出处
《中国计划生育学杂志》
北大核心
2009年第4期227-230,共4页
Chinese Journal of Family Planning
